Expression Of Growth Hormone-releasing Hormone Research Articles

Neonatal malnutrition impacts fibroblast growth factor 21-induced neuron neurite outgrowth and growth hormone-releasing hormone secretion in neonatal mouse brain

Neonatal malnutrition is one of the most common causes of neurological disorders. However, the mechanism of action of the factors associated with neonatal nutrition in the brain remains unclear. In this study, we focused on fibroblast growth factor (FGF) 21 to elucidate the effects of malnutrition on the neonatal brain. FGF21 is an endocrine factor produced by the liver during lactation which is the main source of nutrition during the neonatal period. In this study, malnourishment during nursing mice induced decreased levels of Fgf21 mRNA in the liver and decreased levels of FGF21 in the serum. RNA-seq analysis of neonatal mouse brain tissue revealed that FGF21 controlled the expression of Kalrn-201 in the neonatal mouse brain. Kalrn-201 is a transcript of Kalirin, a Ras homologous guanine nucleotide exchange factor at the synapse. In mouse neurons, FGF21 induced the expression of Kalirin-7 (a Kalirin isoform) by down-regulating Kalrn-201. FGF21-induced Kalirin-7 stimulated neurite outgrowth in Neuro-2a cells. FGF21 also induced Growth hormone-releasing hormone (GHRH) expression in Neuro-2a cells. Kalirin-7 and GHRH expression induced by FGF21 was altered by inhibiting the activity of SH2-containing tyrosine phosphatase (SHP2) which is located downstream of the FGF receptor (FGFR). Additionally, malnourished nursing induced intron retention of the SHP2 gene (Ptpn11), resulting in the alteration of Kalirin-7 and GHRH expression by FGF21 signaling. Ptpn11 intron retention is suggested to be involved in regulating SHP2 activity. Taken together, these results suggest that FGF21 plays a critical role in the induction of neuronal neurite outgrowth and GHRH secretion in the neonatal brain, and this mechanism is regulated by SHP2. Thus, Ptpn11 intron retention induced by malnourished nursing may be involved in SHP2 activity.

Expression of Growth Hormone-Releasing Hormone and Its Receptor Splice Variants in Primary Human Endometrial Carcinomas: Novel Therapeutic Approaches.

Antagonists of growth hormone-releasing hormone (GHRH) inhibit the growth of various tumors, including endometrial carcinomas (EC). However, tumoral receptors that mediate the antiproliferative effects of GHRH antagonists in human ECs have not been fully characterized. In this study, we investigated the expression of mRNA for GHRH and splice variants (SVs) of GHRH receptors (GHRH-R) in 39 human ECs and in 7 normal endometrial tissue samples using RT-PCR. Primers designed for the PCR amplification of mRNA for the full length GHRH-R and SVs were utilized. The PCR products were sequenced, and their specificity was confirmed. Nine ECs cancers (23%) expressed mRNA for SV1, three (7.7%) showed SV2 and eight (20.5%) revealed mRNA for SV4. The presence of SVs for GHRH-Rs could not be detected in any of the normal endometrial tissue specimens. The presence of specific, high affinity GHRH-Rs was also demonstrated in EC specimens using radioligand binding studies. Twenty-four of the investigated thirty-nine tumor samples (61.5%) and three of the seven corresponding normal endometrial tissues (42.9%) expressed mRNA for GHRH ligand. Our findings suggest the possible existence of an autocrine loop in EC based on GHRH and its tumoral SV receptors. The antiproliferative effects of GHRH antagonists on EC are likely to be exerted in part by the local SVs and GHRH system.

Effects of Astragalus Extract Mixture HT042 on Circulating IGF-1 Level and Growth Hormone Axis in Rats.

Astragalus extract mixture HT042 is a standardized functional food granted by the Korean FDA for promoting “Children’s Height Growth”. In this study, we determined whether HT042 affects circulatory Insulin-like growth factor-1 (IGF-1) after administration and investigated whether Growth hormone (GH), Growth hormone-releasing hormone receptor (GHRH-R), and Growth hormone secretagogue receptor (GHS-R) mRNAs are expressed in the pituitary, and whether Growth hormone-releasing hormone (GHRH) and Somatostatin (SST) are expressed in the hypothalamus. We also evaluated the growth effect of HT042 on endochondral bone formation. Male Sprague-Dawley rats in the control and HT042 groups were orally administered a single dose of the control and HT042, respectively, and those in the recombinant human GH (rhGH) group were subcutaneously injected with rhGH. Tetracycline was injected intraperitoneally 72 h prior to sacrifice to decide endochondral bone formation. To determine the endocrine or paracrine/autocrine mechanism, we evaluated the expression of local BMP-2 and IGF-1, an immunohistochemical study after HT042 administration. It was confirmed that the growth-promoting effect of HT042 can be contributed to the increase in serum IGF-1, which can be stimulated by GH secretion. Administration of HT042 modulated the activity of GHRH-R and GHR-S in the pituitary gland and promoted GH secretion, thereby changing longitudinal growth through GH/IGF-1 mediation. Results for GHRH and SST expression demonstrated that the hypothalamus can be influenced and mediated by HT042 through a complex neuroendocrine regulatory system. In addition, it was confirmed by oral administration for 10 days that HT042 increased bone formation in cartilage, which is important for height growth. The effect of HT042 could be owing to upregulation of local Bone morphogenetic protein-2 (BMP-2) and IGF-1 expression in the growth plate, which could be regarded as a GH-dependent autocrine/paracrine pathway, as well as circulatory IGF-1.

Ghrelin Gene Deletion Alters Pulsatile Growth Hormone Secretion in Adult Female Mice.

Using preproghrelin-deficient mice (Ghrl-/-), we previously observed that preproghrelin modulates pulsatile growth hormone (GH) secretion in post-pubertal male mice. However, the role of ghrelin and its derived peptides in the regulation of growth parameters or feeding in females is unknown. We measured pulsatile GH secretion, growth, metabolic parameters and feeding behavior in adult Ghrl-/- and Ghrl+/+ male and female mice. We also assessed GH release from pituitary explants and hypothalamic growth hormone-releasing hormone (GHRH) expression and immunoreactivity. Body weight and body fat mass, linear growth, spontaneous food intake and food intake following a 48-h fast, GH pituitary contents and GH release from pituitary explants ex vivo, fasting glucose and glucose tolerance were not different among adult Ghrl-/- and Ghrl+/+ male or female mice. In vivo, pulsatile GH secretion was decreased, while approximate entropy, that quantified orderliness of secretion, was increased in adult Ghrl-/- females only, defining more irregular GH pattern. The number of neurons immunoreactive for GHRH visualized in the hypothalamic arcuate nucleus was increased in adult Ghrl-/- females, as compared to Ghrl+/+ females, whereas the expression of GHRH was not different amongst groups. Thus, these results point to sex-specific effects of preproghrelin gene deletion on pulsatile GH secretion, but not feeding, growth or metabolic parameters, in adult mice.

GHRH expression plasmid improves osteoporosis and skin damage in aged mice

The hormone secretion of GHRH-GH-IGF-1 axis in animals was decreased as aging. These hormones play an important role in maintaining bone mass and bone structure, and also affect the normal structure and function of the skin. We used plasmid-based technology to deliver growth hormone releasing hormone (GHRH) to elderly mice. In the current study, 80 and 120 μg/kg pVAX-GHRH plasmid expression plasmid were injected into old mice, the serum GHRH and insulin-like growth factor-1(IGF-1) content were increased within three weeks (P < 0.05). In the groups of 80 and 120 μg/kg plasmid, the content of procollagen type I N-terminal pro-peptide (PINP) in the serum was increased(P < 0.05), and the content of C-terminal telopeptides of type I collagen (CTX-1) in the serum was reduced significantly (P < 0.05). Furthermore, the expression of osteoprotegerin (OPG) and osteocalcin (OCN) in the femur also was increased(P < 0.05). The bone mineral density(BMD)、trabecular bone volume (BV/TV) and trabecular number(Tb.N) of mouse femur were increased significantly (P < 0.05) and trabecular separation(Tb.Sp) was decreased(P < 0.05). There were more trabecular bones in the bone marrow cavity and the trabecular bones are thicker in the groups of 80 and 120 μg/kg plasmid relative to control. The superoxide dismutase (SOD) content in the skin was increased(P < 0.05), and the malondialdehyde (MDA) content was reduced significantly (P < 0.05). Meanwhile, the skin moisture content also increased significantly(P < 0.05). Moreover, the expression of matrix metalloproteinase 3(MMP3) and matrix metalloproteinase 9(MMP9) was decreased in the skin(P < 0.05). The thickness of the dermis and epidermis of the skin had increased significantly(P < 0.05). Skin structure is more dense and complete in the two groups. These results indicate that 80 and 120 μg/kg plasmid-mediated GHRH supplementation can improve osteoporosis and skin aging in aged mice.

Lack of Brain Insulin Receptor Substrate-1 Causes Growth Retardation, With Decreased Expression of Growth Hormone-Releasing Hormone in the Hypothalamus.

Insulin receptor substrate-1 (Irs1) is one of the major substrates for insulin receptor and insulin-like growth factor-1 (IGF-1) receptor tyrosine kinases. Systemic Irs1-deficient mice show growth retardation, with resistance to insulin and IGF-1, although the underlying mechanisms remain poorly understood. For this study, we generated mice with brain-specific deletion of Irs1 (NIrs1KO mice). The NIrs1KO mice exhibited lower body weights, shorter bodies and bone lengths, and decreased bone density. Moreover, the NIrs1KO mice exhibited increased insulin sensitivity and glucose utilization in the skeletal muscle. Although the ability of the pituitary to secrete growth hormone (GH) remained intact, the amount of hypothalamic growth hormone-releasing hormone (GHRH) was significantly decreased and, accordingly, the pituitary GH mRNA expression levels were impaired in these mice. Plasma GH and IGF-1 levels were also lower in the NIrs1KO mice. The expression levels of GHRH protein in the median eminence, where Irs1 antibody staining is observed, were markedly decreased in the NIrs1KO mice. In vitro, neurite elongation after IGF-1 stimulation was significantly impaired by Irs1 downregulation in the cultured N-38 hypothalamic neurons. In conclusion, brain Irs1 plays important roles in the regulation of neurite outgrowth of GHRH neurons, somatic growth, and glucose homeostasis.

Increased Expression of Growth Hormone–Releasing Hormone in Fibrinous Inflammation of Proliferative Diabetic Retinopathy

To investigate the involvement of growth hormone-releasing hormone (GHRH) - growth hormone (GH) signaling in pathogenesis of proliferative diabetic retinopathy (PDR). Experimental laboratory study. Vitreous humor, aqueous humor, and serum were obtained from 36 eyes of 36 patients with or without type 2 diabetes from 2017 to 2019. For histologic examination, 6 fibrovascular membranes were excised from eyes with active PDR. Three fibrovascular membranes were excised from nondiabetic patients with proliferative vitreoretinopathy (PVR) as controls. In PDR, the fibrovascular tissues consisted of a mature region containing fibrocytes, and an immature region populated by abundant polymorphonuclear leukocytes in a fibrinogen meshwork. Clusters of leukocytes were found adhering to the vascular walls. In PVR, no fibrinogen and polymorphonuclear leukocyte was observed in the fibrovascular membranes. The levels of GHRH and GH in PDR were significantly increased (P < .001), with 1.8-fold and 72.8-fold in vitreous humor, and 2-fold and 4.9-fold in aqueous humor, respectively, when compared with corresponding levels in controls. No significant difference was detected for insulin-like growth factor-1. Immunohistochemistry showed intense expression of GHRH and its receptor GHRH-R in polymorphonuclear leukocytes, vascular endothelial cells, and fibrocytes in fibrovascular membranes of PDR. GHRH staining was not detectable in infiltrating cells within the fibrovascular membrane of PVR. These findings reveal a possible involvement of GHRH/GHRH-R in fibrinous inflammation that might contribute to the formation of fibrovascular membrane in PDR through mediating activities of leukocytes, vascular endothelial cells, and fibrocytes. Targeting GHRH/GHRH-R may be considered as a potential therapeutic approach for the treatment of PDR.

Expression of Growth and Hunger Related Genes and Physio-Biochemical Responses in Labeo rohita (Hamilton, 1822) Fed with Lysine and Betaine.

The growth promoting effect of lysine and betaine as well as the expression of candidate genes reflecting their efficacy, such as ghrelin, leptin, Growth Hormone Secretagogue Receptor (GHS-R), Insulin like Growth Factor (IGF- 1) and Growth Hormone Releasing Hormone (GHRH) was examined in Labeo rohita fingerlings. One hundred eighty healthy juveniles from a homologous population were randomly distributed to 15 rectangular tanks of 150 litres capacity. The experiment was carried out for 60 days with five treatment groups consisting T1 (0.25% Betaine), T2 (0.5% Betaine), T3 (0.75% Lysine) and T4 (1.5% Lysine) and control group. The experiment was carried out for 60 days with five treatment groups consisting T1 (0.25% Betaine), T2 (0.5% Betaine), T3 (0.75% Lysine) and T4 (1.5% Lysine) and control group. At the end of trial, the growth parameters such as weight gain, SGR, PER were estimated from the weight of the triplicate groups. The digestive, metabolic and antioxidant enzymes were analysed using spectrophotometric methods. The intestine, brain and liver were sampled from the treatments and expression of different genes ghrelin, leptin, GHSR, IGF-1 and GHRH was also performed by realtime PCR. A significant (P<0.05) increase in weight gain, SGR, PER and lowest FCR was found in T4 group which was significantly (p < 0.05) different from other experimental groups. The highest mRNA expression levels of expression were found in T4 group which was similar to that of ghrelin gene mRNA of T2 group. The significantly (p<0.05) highest GHSR, GHRH and IGF-1 gene expression levels were found in T4 treatment group compared to other groups. The present study reveals that the lysine and betaine stimulate growth and expression of ghrelin GHRH, GHS-R and IGF-1 genes. The increase of IGF-I mRNA expression with lysine and betaine supplementation revealed that these compounds act as growth modulators. However, lysine was found to be a more potent modulator of growth compared to betaine.

Growth hormone-releasing hormone receptor mediates cytokine production in ciliary and iris epithelial cells during LPS-induced ocular inflammation

The receptor for growth hormone-releasing hormone (GHRH-R) has been shown to upregulate specifically in the ciliary and iris epithelial cells and infiltrating cells in the aqueous humor in a rat model of acute anterior uveitis. Treatment with GHRHR-R antagonist alleviates significantly these inflammatory responses. Herein we investigated whether the ciliary and iris epithelial cells can respond directly to lipopolysaccharide (LPS) without the influences of circulating leukocytes to produce inflammatory mediators through a GHRH-R mediated mechanism. In explant cultures of rat ciliary body and iris, LPS caused a substantial increase of GHRH-R in 24 h. Immunohistochemistry showed a localization of TLR4, the receptor for LPS, and an elevated expression of IL-6 and IL-1β in ciliary and iris epithelial cells after LPS treatment. LPS also elevated the level of IL-1β, IL-6, and iNOS and increased secretion of IL-1β and IL-6 from the explants. The GHRH-R antagonist, MIA-602, suppressed the elevated expression of IL-1β and IL-6, and reduced the release of IL-6. Such effects were not seen for the GHRHR agonist, MR-409. When co-cultured with leukocytes, expression of GHRH-R in the ocular explants was further enhanced during LPS treatment. Our results demonstrate a direct action of LPS on ciliary and iris epithelial cells to produce pro-inflammatory factors through a GHRH-R mediated mechanism, and suggest a role of these epithelial cells, in addition to the resident antigen presenting cells, in immune surveillance of the eye. Infiltrating leukocytes may enhance these inflammatory responses by regulating GHRH-R in ciliary and iris epithelial cells, in addition to their functions of synthesizing proinflammatory cytokines.

Peripheral activities of growth hormone-releasing hormone (GHRH)

Growth hormone (GH)-releasing hormone (GHRH) is produced by the hypothalamus and stimulates GH synthesis and release in the anterior pituitary gland. In addition to its endocrine role, GHRH exerts a wide range of extrapituitary effects which include stimulation of cell proliferation, survival and differentiation, and inhibition of apoptosis. Accordingly, expression of GHRH, as well as the receptor GHRH-R and its splice variants, has been demonstrated in different peripheral tissues and cell types. Among the direct peripheral activities, GHRH regulates pancreatic islet and β-cell survival and function and endometrial cell proliferation, promotes cardioprotection and wound healing, influences the immune and reproductive systems, reduces inflammation, indirectly increases lifespan and adiposity and acts on skeletal muscle cells to inhibit cell death and atrophy. Therefore, it is becoming increasingly clear that GHRH exerts important extrapituitary functions, suggesting potential therapeutic use of the peptide and its analogs in a wide range of medical settings.

A new approach to the treatment of acute myeloid leukaemia targeting the receptor for growth hormone-releasing hormone.

Growth hormone-releasing hormone (GHRH) is secreted by the hypothalamus and acts on the pituitary gland to stimulate the release of growth hormone (GH). GHRH can also be produced by human cancers, in which it functions as an autocrine/paracrine growth factor. We have previously shown that synthetic antagonistic analogues of GHRH are able to successfully suppress the growth of 60 different human cancer cell lines representing over 20 cancers. Nevertheless, the expression of GHRH and its receptors in leukaemias has never been examined. Our study demonstrates the presence of GHRH receptor (GHRH-R) on 3 of 4 human acute myeloid leukaemia (AML) cell lines-K-562, THP-1, and KG-1a-and significant inhibition of proliferation of these three cell lines invitro following incubation with the GHRH antagonist MIA-602. We further show that this inhibition of proliferation is associated with the upregulation of pro-apoptotic genes and inhibition of Akt signalling in leukaemic cells. Treatment with MIA-602 of mice bearing xenografts of these human AML cell lines drastically reduced tumour growth. The expression of GHRH-R was further confirmed in 9 of 9 samples from patients with AML. These findings offer a new therapeutic approach to this malignancy and suggest a possible role of GHRH-R signalling in the pathology of AML.

Interactions between GHRH and GABAARs in the brains of patients with epilepsy and in animal models of epilepsy

Growth hormone releasing hormone (GHRH) has recently been shown to increase the level of γ-aminobutyric acid (GABA) and activate GABA receptors (GABARs) in the cerebral cortex. GABA is an inhibitory neurotransmitter that can inhibit seizures. Does GHRH enhance the inhibitory effect of GABA to prevent epilepsy by increasing the GABA level and activating GABARs? In this study, patients with epilepsy and C57/BL6 mice with epilepsy induced by kainic acid (KA) or pentylenetetrazol (PTZ) served as the research subjects. Western blots were used to observe the differences in GHRH expression between the normal group and the epilepsy group, immunofluorescence was performed to explore the localization of GHRH in the brain, and coimmunoprecipitation was used to observe the interaction between GHRH and GABARs. GHRH expression was significantly increased in both patients with temporal lobe epilepsy (TLE) and in two mouse models induced by KA or PTZ compared with that in the normal groups (P < 0.05 or P < 0.01). GHRH was expressed in neurons in both humans and mice. Additionally, GHRH co-localized with presynaptic and postsynaptic sites of inhibitory neurons. Coimmunoprecipitation confirmed that GHRH interacted with GABAAα1 and GABAAβ2 + 3. GHRH may play an important role in inhibiting seizures by activating GABAARs.

Inhibitory Effects of Antagonists of Growth Hormone-Releasing Hormone (GHRH) in Thyroid Cancer.

Growth hormone-releasing hormone (GHRH) is a peptide hormone secreted by the hypothalamus that regulates the synthesis and secretion of growth hormone (GH) in the pituitary. The extra-hypothalamic GHRH and its cognate receptors (GHRHR and splice variants) play a mitogenic role by stimulating cell proliferation and preventing apoptotic cell death. It is well established that GHRH antagonists inhibit the growth, tumorigenicity, and metastasis of various human malignancies. In this work, we studied the effect of two new GHRH antagonists, MIA602 and MIA690, on thyroid cancer. We studied the effect of MIA602 and MIA690 on thyroid cancer in vitro, using human thyroid cancer cell lines, and in vivo, using chicken embryo chorioallantoic membrane (CAM) assays. We found that mRNA for GHRH and GHRH receptor is expressed in thyroid cell lines and in samples of thyroid tumors. Immunohistochemistry confirmed the expression of GHRHR protein in specimens of thyroid tumor. We observed that GHRH antagonists inhibited the growth and increased apoptosis of thyroid cancer cells. In vivo, the antagonists inhibited growth and angiogenesis of engrafted thyroid tumors. Our results suggest that GHRH expression may play a role in growth of thyroid cancer and that GHRH antagonists can be a therapeutic option for thyroid cancer patients.

Growth hormone-releasing hormone is produced by adipocytes and regulates lipolysis through growth hormone receptor.

Growth hormone-releasing hormone (GHRH) has a crucial role in growth hormone (GH) secretion, but little is known about its production by adipocytes and its involvement in adipocyte metabolism. To determine whether GHRH and its receptor (GHRH-R) are present in human adipocytes and to study their levels in obesity. Also, to analyze the effects of GHRH on human adipocyte differentiation and lipolysis. GHRH/GHRH-R and GH/GH-R mRNA expression levels were analyzed in human mature adipocytes from non-obese and morbidly obese subjects. Human mesenchymal stem cells (HMSC) were differentiated to adipocytes with GHRH (10-14-10-8 M). Adipocyte differentiation, lipolysis and gene expression were measured and the effect of GH-R silencing was determined. Mature adipocytes from morbidly obese subjects showed a higher expression of GHRH and GH-R, and a lower expression of GHRH-R and GH than non-obese subjects (P<0.05). A total of 10-14-10-10 M GHRH induced an inhibition of lipid accumulation and PPAR-γ expression (P<0.05), and an increase in glycerol release and HSL expression (P<0.05) in human differentiated adipocytes. A total of 10-12-10-8 M GHRH decreased GHRH-R expression in human differentiated adipocytes (P<0.05). A total of 10-10-10-8 M GHRH increased GH and GH-R expression in human differentiated adipocytes (P<0.05). The effects of GHRH at 10-10 M on adipocyte differentiation and lipolysis were blocked when GH-R expression was silenced. GHRH and GHRH-R are expressed in human adipocytes and are negatively associated. GHRH at low doses may exert an anti-obesity effect by inhibiting HMSC differentiation in adipocytes and by increasing adipocyte lipolysis in an autocrine or paracrine pathway. These effects are mediated by GH and GH-R.

The expression of GHRH and its receptors in breast carcinomas with apocrine differentiation—further evidence of the presence of a GHRH pathway in these tumors

Apocrine breast carcinomas were evaluated for the expression of components of the growth hormone-releasing hormone (GHRH) autocrine/paracrine pathway: GHRH and its receptors (GHRH-R), as mammary apocrine carcinomas and epithelium seemed to be uniformly positive for GHRH-R in a pilot study. The apocrine phenotype was determined on the basis of hematoxylin-eosin morphology and a congruent immunohistochemical profile (estrogen receptor negativity, androgen receptor and gross cystic disease fluid protein-15 positivity). Thirty-five formalin-fixed, paraffin-embedded apocrine breast cancers in tissue microarrays and 24 cases using whole-tissue sections were evaluated for GHRH-R and GHRH expression by immunohistochemistry using polyclonal antibodies raised against various domains of GHRH-R and one polyclonal antibody specific for GHRH. GHRH-R positivity was detected in the overwhelming majority (ranging from 90% to 100%) of apocrine breast carcinomas with all but one of the antibodies applied. The expression was usually diffuse with only isolated cases showing positivity in less than 50% of tumor cells. With the PA5-33583 antibody, GHRH-R positivity was seen only in 73% of the cases in at least 50% of the tumor cells. GHRH expression was also present in all but one case tested, with more than 50% of the cells expressing it in 30/34 cases. These results support a high rate of GHRH-R and GHRH expression in apocrine breast carcinomas. Whether these findings can be exploited for the targeted treatment of apocrine breast carcinomas with GHRH antagonists requires further study.

MP17-03 INFLAMMATION-INDUCED PROSTATIC ENLARGEMENT AND PROLIFERATION OF PROSTATE EPITHELIAL CELLS IS REDUCED BY GROWTH HORMONE-RELEASING HORMONE (GHRH) ANTAGONISTS THROUGH THE INHIBITION OF EPITHELIAL-TO-MESENCHYMAL TRANSITION

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2017MP17-03 INFLAMMATION-INDUCED PROSTATIC ENLARGEMENT AND PROLIFERATION OF PROSTATE EPITHELIAL CELLS IS REDUCED BY GROWTH HORMONE-RELEASING HORMONE (GHRH) ANTAGONISTS THROUGH THE INHIBITION OF EPITHELIAL-TO-MESENCHYMAL TRANSITION Petra Popovics, Andrew Schally, Luis Salgueiro, Krisztina Kovacs, and Ferenc Rick Petra PopovicsPetra Popovics More articles by this author , Andrew SchallyAndrew Schally More articles by this author , Luis SalgueiroLuis Salgueiro More articles by this author , Krisztina KovacsKrisztina Kovacs More articles by this author , and Ferenc RickFerenc Rick More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.592AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The pathogenesis of benign prostatic hyperplasia (BPH) has been associated with various factors including hormonal imbalance, inflammation-induced cell proliferation and epithelial-to-mesenchymal transition. We have previously demonstrated that prostatic GHRH and its receptors are upregulated in a rat testosterone-induced BPH model and GHRH antagonists suppress the levels of proinflammatory cytokines. Based on these findings, we investigated the role of GHRH in inflammation-induced proliferation of prostate epithelial cells in vitro and prostate enlargement in experimental autoimmune prostatitis. METHODS Autoimmune inflammation in the prostates of Balb/c mice was induced by subcutaneous injections of rat male tissue homogenate. Changes in prostate volume were measured with the VEVO® 1100 ultrasound imaging system. Human BPH-1 and primary prostate epithelial cells were used in matrigel-embedded 3D cultures and average sphere diameters were evaluated. Chronic inflammation was mimicked by treating cells with THP-1 macrophage-conditioned medium or Il-17A, whereas EMT was triggered with TGF-β1 or TGF-β2 peptides. The role of secreted GHRH in inflammation-induced proliferation was determined by using GHRH antagonists developed in our lab. Changes in the protein levels were determined by western blot. RESULTS Experimental autoimmune prostatitis increased the volume of the ventral prostate by 92% at week 8 compared to control (p<0.001). A 1-month daily treatment with GHRH antagonists caused a significant, 48% reduction in prostate volume. Macrophage-conditioned medium induced a 26% increase (p<0.001) in the average diameter of cells and elevation in the expression of mesenchymal markers. The mRNA and protein expression of GHRH were significantly increased by macrophage-conditioned medium. GHRH antagonist reduced inflammation- and TGF-β2-induced increase in diameter by 64% (p<0.01) and by 67% (p<0.001), respectively, and reduced the expression of n-cadherin. The stimulatory effect of TGF-β2 was abolished when GHRH receptor expression was silenced by stable transfection of shRNA. IL-17A stimulation of the growth of primary epithelial cells were also significantly reduced by GHRH antagonists. CONCLUSIONS Our results indicate that GHRH is a key factor in prostatic inflammation-induced prostate enlargement and EMT and suggest that GHRH antagonists have beneficial effects in chronic prostatitis and BPH. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e212 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Petra Popovics More articles by this author Andrew Schally More articles by this author Luis Salgueiro More articles by this author Krisztina Kovacs More articles by this author Ferenc Rick More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

670 Growth hormone-induced signaling as a novel, intrafollicular neuroendocrine control of human hair growth

Growth hormone (GH) and activation of its receptor (GHR) promote cell growth, proliferation, differentiation and stem cell activation either directly or via the induction of IGF-1. While some clinical case reports suggest that human hair follicles (HFs) may be GH-responsive, it is unknown whether GH exerts any functions in human HF biology. To explore this we charted the expression of GH, GHR and GHRH (growth hormone releasing hormone), the hypothalamic neurohormone that stimulates pituitary GH expression and release, in human scalp HFs. So far, we have detected GH, GHR and GHRH protein expression in the ORS of human anagen VI HFs as well as in the sebaceous gland epithelium. qRT-PCR analysis revealed that human scalp HFs also transcribe GHR mRNA, supporting that human HFs are an extrapituitary source of GH production. qRT-PCR analysis showed that GHR transcription decreases during HF regression (catagen HFs). Interestingly, the level of GHR transcription was inversely correlated with that of the GH-inhibiting hormone, somatostatin (SST), whose expression increased in catagen HFs. Next, organ-cultured human scalp HFs were stimulated with recombinant hGH (rhGH, 50-100 ng/ml) or recombinant GH-binding protein (rGHBP), which can exert both GH-antagonistic and agonistic effects. hGH and GHBP both promoted hair shaft elongation and tended to inhibit catagen development ex vivo. qRT-PCR analysis showed that hGH (100 ng/ml) and GHBP (100 ng/ml) induced an increase in HF mRNA levels of JAK2, the downstream target after GHR activation, and of SST. While we are still investigating how GHR silencing in human HFs ex vivo impacts on standard hair biology read-out parameters, our pilot study already suggests that GHR-mediated signaling by intrafollicularly generated GH is as a major novel neuroendocrine regulator of human hair growth.

MP44-11 GROWTH HORMONE-RELEASING HORMONE (GHRH) ANTAGONISTS REDUCE INFLAMMATION- AND TRANSFORMING GROWTH FACTOR (TGF)-?2-INDUCED PROLIFERATION OF HUMAN BPH-1 PROSTATE EPITHELIAL CELLS GROWN IN 3D CULTURE

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2016MP44-11 GROWTH HORMONE-RELEASING HORMONE (GHRH) ANTAGONISTS REDUCE INFLAMMATION- AND TRANSFORMING GROWTH FACTOR (TGF)-?2-INDUCED PROLIFERATION OF HUMAN BPH-1 PROSTATE EPITHELIAL CELLS GROWN IN 3D CULTURE Petra Popovics, Andrew V. Schally, Roberto Perez, Rhenzi Cai, and Ferenc G. Rick Petra PopovicsPetra Popovics More articles by this author , Andrew V. SchallyAndrew V. Schally More articles by this author , Roberto PerezRoberto Perez More articles by this author , Rhenzi CaiRhenzi Cai More articles by this author , and Ferenc G. RickFerenc G. Rick More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.267AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Benign prostatic hyperplasia (BPH) is frequently associated with chronic inflammation. Epithelial-to-mesenchymal transition (EMT) has been recently indicated as a major factor in the pathogenesis of BPH. Evidence also suggests that inflammatory cytokines may induce BPH. Hypothalamic neuropeptides, GHRH, luteinizing hormone-releasing hormone (LHRH) and their receptors are expressed in experimental models of BPH in which antagonists of GHRH and LHRH have been shown to suppress the levels of proinflammatory cytokines and alter the expression of genes related to EMT. These findings imply a role of GHRH/LHRH in the development of prostatitis, however, this has not been investigated to date. METHODS Human BPH-1 prostatic cell line was used to generate matrigel-embedded 3D cultures in which average sphere diameters were evaluated. Chronic inflammation was induced by treating cells with THP-1 macrophage-conditioned medium whereas EMT was triggered with TGF-ß1 or TGF-ß2 peptides. The role of secreted LHRH and GHRH in inflammation-induced proliferation was determined by using the LHRH antagonist, degarelix, and a specific GHRH antagonist developed in our lab. The presence of receptors for GHRH and LHRH was confirmed in 3D cultures by immunocytochemistry and quantitative RT-PCR. RESULTS BPH-1-derived spherical structures expressed cell membrane receptors for LHRH and GHRH. Macrophage-conditioned medium induced a 26% increase (P<0.001) in the average diameter of cells and a 2-fold elevation in the expression of the EMT markers N-cadherin (p<0.01) and Snail (p<0.001) as detected by qPCR. The expression of GHRH was also increased 2.7-fold (p<0.001), whereas mRNA levels of LHRH were slightly reduced (ns). GHRH antagonist reduced inflammation-induced increase in diameter by 64% (P<0.01), while no significant change was observed after degarelix treatment. TGF-ß2 increased average sphere diameter by 32%, but this effect was reduced by 67% by the GHRH antagonist (P<0.001). In contrast, TGF-ß1 did not significantly affect sphere diameter. CONCLUSIONS This study identifies GHRH as an important factor in inflammation-induced proliferation and TGF-ß2-triggered EMT of prostate epithelial cells and suggests the merit of further investigation to elucidate the effects of GHRH antagonists in the reduction of inflammation-induced BPH. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e602-e603 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Petra Popovics More articles by this author Andrew V. Schally More articles by this author Roberto Perez More articles by this author Rhenzi Cai More articles by this author Ferenc G. Rick More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Peripheral activities of growth hormone-releasing hormone.

Growth hormone (GH)-releasing hormone (GHRH) is produced by the hypothalamus and stimulates GH synthesis and release in the anterior pituitary gland. In addition to its endocrine role, GHRH exerts a wide range of extrapituitary effects which include stimulation of cell proliferation, survival and differentiation, and inhibition of apoptosis. Accordingly, expression of GHRH, as well as the receptor GHRH-R and its splice variants, has been demonstrated in different peripheral tissues and cell types. Among the direct peripheral activities, GHRH regulates pancreatic islet and β-cell survival and function and endometrial cell proliferation, promotes cardioprotection and wound healing, influences the immune and reproductive systems, reduces inflammation, indirectly increases lifespan and adiposity and acts on skeletal muscle cells to inhibit cell death and atrophy. Therefore, it is becoming increasingly clear that GHRH exerts important extrapituitary functions, suggesting potential therapeutic use of the peptide and its analogs in a wide range of medical settings.

Prognosis in human glioblastoma based on expression of ligand growth hormone-releasing hormone, pituitary-type growth hormone-releasing hormone receptor, its splicing variant receptors, EGF receptor and PTEN genes.

Glioblastoma (GB) is the most frequent brain tumor. Despite recent improvement in therapeutic strategies, the prognosis of GB remains poor. Growth hormone-releasing hormone (GHRH) may act as a growth factor; antagonists of GHRH have been successfully applied for experimental treatment of different types of tumors. The expression profile of GHRH receptor, its main splice variant SV1 and GHRH have not been investigated in human GB tissue samples. We examined the expression of GHRH, full-length pituitary-type GHRH receptor (pGHRHR), its functional splice variant SV1 and non-functional SV2 by RT-PCR in 23 human GB specimens. Epidermal growth factor receptor (EGFR) and phosphatase and tensin homolog gene (PTEN) expression levels were also evaluated by quantitative RT-PCR. Correlations between clinico-pathological parameters and gene expressions were analyzed. Expression of GHRH was found to be positive in 61.9 % of samples. pGHRH receptor was not expressed in our sample set, while SV1 could be detected in 17.4 % and SV2 in 8.6 % of the GB tissues. In 65.2 and 78.3 % of samples, significant EGFR over-expression or PTEN under-representation could be detected, respectively. In 47.8 % of cases, EGFR up-regulation and PTEN down-regulation occurred together. Survival was significantly poorer in tumors lacking GHRH expression. This worse prognosis in GHRH negative group remained significant even if SV1 was also expressed. Our study shows that GHRH and SV1 genes expressed in human GB samples and their expression patterns are associated with poorer prognosis.